Differentiation of homokaryons and heterokaryons of Agaricus bisporus with inter-simple sequence repeat markers

Microbiological Research
Mahmudul Islam Nazrul, Bian YinBing

Abstract

Morphologically different fifteen slow growing single spore isolates (SSIs) were screened from germinated basidiospores of Agaricus bisporus; assumed to be homokaryons, and subjected to growth rate, spawn run, fruiting and inter-simple sequence repeat (ISSR) analysis for confirmation of true homokaryons. The present studies are the first report of fingerprinting on differentiating homo- and hetero-karyotic SSIs using ISSR markers. The patterns were highly polymorphic and very reproducible. Among 40 primers tested, 7 ISSR primers were selected for the analysis of genomic DNA and generated a total of 54 ISSR fragments, sufficient to differentiate the 15 isolates from each other. ISSR fingerprinting detected 46.30% polymorphic loci. All appressed homokaryons carried a subset of ISSR markers found in the heterokaryons, and these were not able to produce a fruiting body. A test of cross-fertility and the following fruiting trial proved that 3 of the 15 SSIs with different ISSR fingerprints were homokaryons. None of the discarded SSIs have any missing bands present in the parental heterokaryotic control; these are heterokaryons derived from non-sister nuclear pairs. It is revealed that homokaryons are probably restricted to the assum...Continue Reading

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